construction of recombinant bacmid dna encoding newcastle disease virus (ndv) fusion protein gene

background and aims: newcastle disease virus (ndv) is one of the major pathogen in poultry. vaccination is intended to control the disease as an effective solution nevertheless this virus is a growing threat to the poultry industry. f gene open reading frame (orf) from ndv is 1650 bp, encoding a protein of 553 amino acids that can induce protective immunity alone. the f glycoprotein on the surface of ndv is important for virus infectivity and pathogenicity. towards protection goal, the full-length of f gene was isolated using specific primers and cloned into the baculovirus derived bacmid shuttle vector to produce recombinant f-protein in insect cells. materials and methods: f gene orf from avirulent strain la sota of ndv was amplified by rt-pcr to produce f cdna. the amplicon was cloned firstly into the t/a cloning vector and then subcloned into the pfastbac dual donor plasmid through ncoi/kpni sites. after the verification of cloning process by pcr and enzymatic digestion analysis, the accuracy of f gene orf in the t/a cloning vector was confirmed by sequencing. finally, f-containing recombinant bacmid was subsequently generated and confirmed by pcr using f specific primers and m13 universal primers. results: results showed that a recombinant baculovirus containing a correct and in framework sequence of newcastle f gene under the control of p10 promoter was constructed. conclusion: the above mentioned f-containing recombinant baculovirus, in addition to its independent application, can be used with other individual recombinant baculoviruses expressing nh and np genes to produce newcastle vlps in insect cell line.